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Identifying Biomarkers of Heterogeneity and Transplantation Efficacy in Retinal Pigment Epithelial Cells [Bulk RNA-seq]

ABSTRACT: Identifying Biomarkers of Heterogeneity and Transplantation Efficacy in Retinal Pigment Epithelial Cells [Bulk RNA-seq]

PROVIDER: PRJNA869448 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21046616_1.fastq.gz	Fastqsanger.gz
	SRR21046616_2.fastq.gz	Fastqsanger.gz
	SRR21046617_1.fastq.gz	Fastqsanger.gz
	SRR21046617_2.fastq.gz	Fastqsanger.gz
	SRR21046618_1.fastq.gz	Fastqsanger.gz

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Identifying biomarkers of heterogeneity and transplantation efficacy in retinal pigment epithelial cells

Project description:Identifying biomarkers of heterogeneity and transplantation efficacy in retinal pigment epithelial cells

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Identifying biomarkers of heterogeneity and transplantation efficacy in retinal pigment epithelial cells

Project description:Transplantation of retinal pigment epithelial (RPE) cells holds great promise for patients with retinal degenerative diseases, such as age-related macular degeneration. In-depth characterization of RPE cell product identity and critical quality attributes are needed to enhance efficacy and safety of replacement therapy strategies. Here, we characterized an adult RPE stem cell-derived (RPESC-RPE) cell product using bulk and single-cell RNA sequencing (scRNA-seq), assessing functional cell integration in vitro into a mature RPE monolayer and in vivo efficacy by vision rescue in the Royal College of Surgeons rats. scRNA-seq revealed several distinct subpopulations in the RPESC-RPE product, some with progenitor markers. We identified RPE clusters expressing genes associated with in vivo efficacy and increased cell integration capability. Gene expression analysis revealed lncRNA (TREX) as a predictive marker of in vivo efficacy. TREX knockdown decreased cell integration while overexpression increased integration in vitro and improved vision rescue in the RCS rats.

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Identifying Biomarkers of Heterogeneity and Transplantation Efficacy in Retinal Pigment Epithelial Cells [scRNA-seq]

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Project description:Transplantation of retinal pigment epithelial (RPE) cells holds great promise for patients with retinal degenerative diseases such as age-related macular degeneration. In-depth characterization of RPE cell product identity and critical quality attributes are needed to enhance efficacy and safety of replacement therapy strategies. Here we characterized an adult RPE stem cell-derived (RPESC-RPE) cell product using bulk and single cell RNA sequencing (sc-RNA-seq), assessing functional cell integration in vitro into a mature RPE monolayer and in vivo efficacy by vision rescue in the Royal College of Surgeons rats. scRNA-seq revealed several distinct subpopulations in the RPESC-RPE product, some with progenitor markers. We identified RPE clusters expressing genes associated with in vivo efficacy and increased cell integration capability. Gene expression analysis revealed a lncRNA (TREX) as a predictive marker of in vivo efficacy. TREX knockdown decreased cell integration while overexpression increased integration in vitro and improved vision rescue in the RCS rats.

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Identifying Biomarkers of Heterogeneity and Transplantation Efficacy in Retinal Pigment Epithelial Cells [Bulk RNA-seq]

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Project description:Retinal Pigment Epithelium (RPE) are crucial for maintaining retinal homeostasis and the visual cycle. Currently, there are no effective treatments for blinding retinal diseases caused by RPE cell degeneration. Gene therapy presents a promising approach, but no gene therapy vectors specifically targeting RPE cells are available. In this study, we utilized an AAV2 random mutation library and high-throughput sequencing to screen AAV vectors （AAV206）capable of specifically targeting RPE cells. Intravitreal injection of the AAV206 vector in mice demonstrated that AAV206 specifically infected RPE cells, whereas AAV2 primarily infects the inner retina. Furthermore, AAV206 exhibited lower immunogenicity and toxicity compared to AAV2. Overexpression of sFLT-1 through AAV206 effectively inhibited the size and leakage of choroidal neovascularization. In summary, AAV206 is an ideal vector for gene therapy targeting RPE cells.

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